Fig. 5: HPF1 stimulates initiation in PARP1. | Nature Communications

Fig. 5: HPF1 stimulates initiation in PARP1.

From: HPF1 dynamically controls the PARP1/2 balance between initiating and elongating ADP-ribose modifications

Fig. 5

a PARP1 (1 μM) was incubated without or with HPF1 (0.1 μM) for 10 min at RT in the presence of DNA (1 μM). 1 mM NAD+ was added for various time points as indicated, and reactions were quenched with 500 μM talazoparib. PARG (1 μM) was added and incubated for 1 h at RT. Reactions were resolved by 12% SDS-PAGE and a western blot was performed using a mono-ADP-ribose-binding reagent. The bands corresponding to mono-ADP-ribosylated PARP1 were quantified using ImageJ and plotted as intensity over time. b Reactions were performed as in panel a without HPF1 and comparing PARP1 WT to PARP1 H826E. Reactions were resolved by 7.5% SDS-PAGE and a western blot was performed using a mono-ADP-ribose binding reagent. For 5a and b, a representative image from three independent experiments is shown. Numbers on the left side of the blots represent molecular weight marker locations (values in kDa). Source data are provided as a Source Data file.

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